...Resistance Activity PI 61422 dCAS9-VP64_GFP dCAS9 (D10A, H840A) none Expresses dCAS9-VP64 activator with 2A GFP... dCAS-VP64_Blast dCAS9 (D10A, N863A) Blasticidin 3rd generation lenti vector encoding dCAS9-VP64 with ...with 2A Blast resistance marker (EF1a-NLS-dCas9(N863)-VP64-2A-Blast-WPRE) Zhang 61426 lenti MS2-P65-HSF1_...56,762 unique sgRNAs. This backbone does not contain dCas9. 56,762 unique sgRNAs targeting 18,885 human genes...Human CRISPR inhibition library Dolcetto in XPR_050 (VP64) Set A Human CRISPR inhibition library in backbone...non-targeting controls. This backbone does not contain dCas9. 57,050 unique sgRNAs targeting 18,901 human genes...Human CRISPR inhibition library Dolcetto in XPR_050 (VP64) Set B Human CRISPR inhibition library in backbone...

...46912 pMSCV-LTR-dCas9-VP64-BFP Human expression vector containing MSCV LTR promoter, dCas9 that is fused...to 2x NLS, VP64 and tagBFP 51023 pSLQ1658-dCas9-EGFP Human expression vector containing dCas9 that is fused...constructs 46910 pHR-SFFV-dCas9-BFP Human expression vector containing SFFV promoter, dCas9 that is fused to 2x... 46911 pHR-SFFV-dCas9-BFP-KRAB Human expression vector containing SFFV promoter, dCas9 that is fused to...46913 pMSCV-LTR-dCas9-p65AD-BFP Human expression vector containing MSCV LTR promoter, dCas9 that is fused... CXCR4 gene 46920 pTDH3-dCas9 Yeast CEN/ARS vector (Leu2) that contains dCas9 fused to NLS controlled ... promoter 46921 pTDH3-dCas9-Mxi1 Yeast CEN/ARS vector (Leu2) that contains dCas9 fused to N 51025 pSLQ1661...

...-pCR8-dCas9VP48 : dCas9VP48 on Gateway donor vector pCR8/GW/TOPO. 48218 pAC84-pCR8-dCas9 : dCas9 on Gateway...components, the dCas9 activator protein and the single guide RNA (sgRNA). While the dCas9 protein provides... 3x minimal VP16 domains, VP64 with 4, VP96 with 6, and VP160 with 10. dCas9 activators and sgRNAs can...below: Table 1. dCas9 Activators sgRNA Dual Expression ID Plasmid 48236 pAC2-dual-dCas9VP48-sgExpression ...construct expressing both dCas9VP64 and sgRNA from separate promoters. Table 2. pmax dCas9 Activator Expression... dCas9VP160-2A-puro (puro-selectable) on pmax expression vecor. Table 3. dCas9 Activators Gateway Donors...can be separately transfected (pmax dCas9 activators [48223~48227] + sgRNA expression vector or the linear...

...for transient transfection : 61422 : Expresses dCAS9-VP64 activator with 2A GFP 61423 : Expresses the MS2...production : 61425 : lentiviral vector encoding dCAS9-VP64 with 2A Blast resistance marker 61426 : lentiviral...three components: A nucleolytically inactive Cas9-VP64 fusion A sgRNA incorporating two MS2 RNA aptamers...

...permanently modifying the genome dCas9-repressor (such as dCas9-KRAB) or dCas9 gRNA(s) targeting promoter elements...permanently modifying the genome dCas9-activator (such as dCas9-VP64) gRNA(s) targeting promoter elements... elements of target gene dCas9-KRAB is more effective than dCas9 alone for mammalian cell lines. Activate...activate or repress a target gene using dCas9-activators or dCas9-repressors, gRNAs should be targeted to...

...Mammalian Lentiviral expression of Cas9, dCas9, or dCas9-VP64 along with 1-4 sgRNAs expressed from independent...separate accessory pack is available for dCas9 and FokI-dCas9. Yamamoto Multiplex CRISPR/Cas9-based genome...Editing Other Applications Purify Tag Visualize dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors...Mammalian U6 yes, nick S. pyogenes Conklin pXFokI-dCas9 60901 Mammalian U6 yes, FokI S. pyogenes Conklin...Mammalian U6 none S. aureus Joung pLV hU6-sgRNA hUbC-dCas9-KRAB-T2a-Puro 71236 Mammalian/Lentiviral hU6 yes...Lentiviral U6 none S. pyogenes Puro Zhang pRS416-dCas9-Mxi1 + TetR + pRPR1(TetO)-NotI-gRNA 73796 Yeast ...pyogenes Chen pdCas9-DNMT3A-EGFP 71666 Mammalian U6 yes, methylation S. pyogenes EGFP Zoldos pdCas9-DNMT3A-PuroR...

...achieved by using orthogonal dCas9s (e.g. S. pyogenes dCas9 and S. aureus dCas9) tagged with different fluorescent...human cells. The simplest dCas9-based activators and repressors consist of dCas9 fused directly to a single...imaging of proteins in living cells (Figure 9B) dCas9-VPR - dCas9 fused to several different activation domains...(Figure 9C) SAM activators - co-expression of dCas9-VP64 with a modified scaffold gRNA and additional ...utilizes biotin tagging of dCas9 by fusing a biotin acceptor site to dCas9 and co-expressing BirA biotin... defense CRISPRa CRISPR Activation; using a dCas9 or dCas9-activator with a gRNA to increase transcription...target gene CRISPRi CRISPR Interference; using a dCas9 or dCas9-repressor with a gRNA to repress/decrease transcription...